Antibody Characterization of Dystroglycan and Sarcoglycan

Monoclonal antibodies IIH6 against α-dystroglycan (Ervasti and Campbell, 1991 (link)) and 8D5 against β-dystroglycan (Lim et al., 1995 (link)) were previously characterized. mAbs 20A6 against α-sarcoglycan, 5B1 against β-sarcoglycan, and 21B5 against γ-sarcoglycan were generated in collaboration with L.V.B. Anderson (Newcastle General Hospital, Newcastle upon Tyne, UK). We used a mAb against caveolin-3 (Transduction Laboratories, Lexington, KY). Rabbit polyclonal antibodies against α-sarcoglycan (Roberds et al., 1993a), dystrophin, and utrophin (Ohlendieck et al., 1991a), neuronal nitric oxide synthase (Crosbie et al., 1998 (link)), the α₁ subunit of the dihyrdopyridine receptor (Ohlendieck et al., 1991b), and the laminin α2-chain (Allamand et al., 1997 (link)) were described previously. Two affinity-purified rabbit antibodies (rabbit 208 and 215) were produced against a full-length COOH-terminal fusion protein of γ-sarcoglycan, and against an NH₂-terminal peptide (MMPQEQYTHHRSTMPGAA) of δ-sarcoglycan, respectively. An affinity-purified goat antibody (goat 26) was produced against a NH₂-terminal fusion protein of β-sarcoglycan containing amino acids 1–65. Polyclonal antibodies against α-dystroglycan fusion protein D were affinity-purified from goat 20 (Ibraghimov-Beskrovnaya et al., 1992 (link)). An affinity-purified rabbit antibody (rabbit 235) was produced against a COOH-terminal fusion protein of sarcospan (CFVMWKHRYQVFYVGVGLRSLMASDGQLPKA). Two polyclonal antibodies against ε-sarcoglycan were used. One was previously characterized (Ettinger et al., 1997 (link)) and the other (rabbit 232) was generated against a COOH-terminal peptide of ε-sarcoglycan (PHQTQIPQQQTTGKWYP).